Apoptosis Compound Library

In cultured mammalian cells disruption of ATP citrate lyase, an enzyme that supplies nucleocytoplasmic acetyl CoA. Conversely, reduced expression of acetyl CoA carboxylase 1, a cytosolic enzyme that competes with HATs for nucleocytoplasmic acetyl CoA, caused an increase in bulk histone acetylation. Moreover, a surge in intracellular acetyl CoA during the oxidative phase of yeast metabolic cycles induced the Gcn5p/SAGA-catalysed acetylation of histones at growth genes and acted as a trigger for initiating a cellular growth programme and cell cycle entry. Intracellular acetyl CoA levels have also been implicated in regulation of Na-linked acetylation and apoptosis. Overexpression of Bcl-xL in human cells caused a reduction in cellular acetyl CoA and a concomitant decrease in protein Na-acetylation, which could be restored by increasing acetyl CoA levels by addition of citrate or acetate. High levels of acetyl CoA and Na-acetylation were shown to be associated with increased susceptibility to apoptotic stimuli. Considering the emerging role of intracellular acetyl CoA levels in the regulation of cell growth, differentiation, cell cycle, and apoptosis, we sought to measure changes in the level of this metabolite in vivo during the embryonic development of a vertebrate. Most in vivo measurements of CoA species have been limited to tissues of adult organisms subjected to different conditions, whereas how the levels of CoA species change in a developing organism is largely unknown. In this study we used embryos of Xenopus laevis, a widely used model species for studying cellular processes underlying embryogenesis. In the present study, we have used Xenopus laevis as a model organism to measure changes in whole-embryo levels of CoA and acetyl CoA in vivo during vertebrate embryonic development. As far as we are aware, the only other study that has measured acetyl CoA levels during the early embryonic development is that by Vastag et al, who employed a mass spectrometry based approach to measure changes in 48 common metabolites, including acetyl CoA, during early Xenopus embryonic development. Based on their data they concluded that there is no observable change in acetyl CoA levels between fertilisation and 11 h post-fertilisation. This contradicts our data showing a small but statistically significant increase in acetyl CoA levels between stage 4 and stage 8/9. The discrepancy may be explained by the different approaches used by the two studies for sampling and data analysis. Vastag et al measured metabolites in each of 10– 11 individual embryos, obtained from three different clutches of eggs, at five different stages between fertilisation and stage 9. Data from individual embryos were then analysed and presented separately. This approach was used to identify metabolites whose concentrations change robustly and consistently in every embryo.

At the same time lowering the stress-level and depressive symptoms. It has previously been theorized that therapies such as acupuncture may exert their effects through activation of DNIC. The gold standard in measuring pain sensitivity is by algometry. Recently a simple and handheld algometric device has been designed to asses pressure pain sensitivity. The PPS measure has in patients with IHD been found to be significantly correlated to the major depression inventory score, WHO5’s well-being index as well as to the SF-36 quality of life score. Several studies have evaluated the effect of various stressreducing interventions in patients with IHD and some have shown to improve the prognosis and to reduce the risk of new cardiovascular events. We hypothesize that an intervention built on an increased focus on stress and the ability to perform stress reduction should be beneficial for patients with IHD. In analogy to people with diabetes measuring blood glucose levels, a therapy based on a daily semi-objective stress-measurements based on PPS followed by stress-reducing actions theoretically leads to increased empowerment and may have a positive effect on stressparameters. Acupressure, i.e. applying a continuous pressure for approximately one minute at specific hyperalgesic points at the body, has been shown to reduce both local and spreading pain in chronic low back pain and neck pain syndromes and we have observed that acupressure results in an acute reduction in pain sensitivity and PPS. Thus we hypothesized that the combination of daily selfmeasurements of PPS aiming at increased empowerment followed by acupressure aiming to restore DNIC, together would resolve in a reduction of the following elements of chronic stress: Depressive symptoms measured by MDI, general well-being measured by WHO-5, and physically and mentally QOL measured by SF-36 QOL-score. Aiming to test this hypothesis we performed an observer blinded randomized clinical trial with blinded outcome assessment over 3 month in which the active group measured PPS twice daily followed by acupressure as mandatory action. Our primary end point was changes in MDI. In the present randomized interventional trial we found, that in patients with stable IHD the combination of daily self-measurements of PPS followed by acupressure and reflection on PPS as a surrogate for current stress-level resulted in a modest but statistically significant improvement in PPS, MDI and WHO-5. This beneficial effect of the intervention program was more pronounced in subgroups of patients with higher baseline levels of components of the chronic stress syndrome, such as elevated MDI, as well as the combination of an elevated PPS and CSS. These subgroup analyses, however, were hampered by a limited sample size. The primary endpoint was a reduction in MDI. MDI was chosen as a well validated often used psychometric questionnaire on depressive symptoms.

Around each of the slopes at each baseline HbA1c. This would permit a variety of scenarios such as high baseline HbA1c, negative slope, low standard deviation, etc. However, the trajectories that we found are empirical, reflecting the true reality of our sample, and converged to 6 trajectories, which do not cover all the spectrum of trajectories to, theoretically, enable such an examination. This is a limitation of an observational study, but, a randomized trial where patients with high initial HbA1c levels were treated/not treated to decrease it, would be unethical. We nevertheless, performed, on the full sample, secondary analyses examining the relationships of mean and standard deviation of HbA1c with the cognitive outcomes and found that higher mean and standard deviation in HbA1c measurements over time were associated with lower scores in overall cognitive score and in executive functions, consistent with the trajectories results. Brain imaging was not performed in this study, thus limiting our ability to evaluate the contribution of cerebrovascular abnormalities to the association of trends in glycemic control with cognition. This is particularly relevant both because T2D is a vascular disease, but also because trajectories of HbA1c were not associated with episodic memory, suggesting non AD-related mechanisms. Entry into the DR, rather than time of T2D diagnosis, which was not available to us, is referred to as “baseline”. Although women are slightly under-represented in the study, sex was one of the covariates in the comparison of groups. Israel has a strong family oriented culture, so a major role in grand- parenting was the primary reason of refusal by women to participate in the study. Additional strengths of this study include the large sample, validated T2D diagnosis for each subject, an average of 18 HbA1c measurements permitting investigation of trajectories of HbA1c over time, strong validity for risk factor levels and medical diagnosis, and a thorough cognitive evaluation. Fluorescent reporter proteins derived from the green fluorescent protein and its derivatives have been used to analyze a wide array of cellular processes such as gene regulation, protein localization, and protein interaction. However, GFP-based fluorescent proteins require molecular oxygen for the generation of cyclic tripeptide chromophore, which has been a major drawback in the GFP-based reporter protein applications under conditions with limited oxygen supply. Alternatively, the flavin mononucleotide -based fluorescent proteins can be employed as an in vivo reporter system. Bacterial FbFPs respond to light blue and activate stress-related signaling pathways. Among these FbFPs, Escherichia coli FbFP which consists of 137 amino acids was engineered from the photoactive LOV domain of Bacillus subtilis YtvA, which is a 261-amino acid protein consisting of two functional domains, a core LOV domain and a sulfate transporter.

Only a minority of the oral microbial community can adhere to hard and soft oral tissues, and assembly of the complex oral biofilm is accomplished by subsequent adherence of secondary colonizers. Streptococcus is an early colonizer and Fusobacterium has a propensity for co-aggregation with many genera, forming a bridge between early and late colonizers in the oral biofilm. Thus, on the one hand, the observed decrease in prevalence of Streptococcus and increased abundance of Fusobacterium genera in pre-cancers could reflect the altered surface properties of the cancer cells and stroma, which might no longer support adhesion of streptococci. On the other hand, we can hypothesize that shifts in abundance of these two genera could result in an enhanced pro-inflammatory environment, since Streptococcus species have been reported to attenuate Fusobacterium nucleatum induced pro-inflammatory responses of oral epithelial cells. We also note that Fusobacterium nucleatum grown as a biofilm is capable of invading organotypic cultures, and secondly, that the organism has recently been reported in colon cancers, further supporting a potential role in oral cancer. The oral cavity offers a relatively unique opportunity to screen at risk individuals for cancer, because the lesions can be seen, and as we report here, the shift in the microbiome of the cancer and pre-cancer lesions compared to anatomically matched clinically normal tissue from the same individual can be detected in non-invasively collected swab samples. Saliva is another noninvasively collected oral sample composed largely of bacterial cells, but also shed epithelial and immune cells. A variety of “omics” biomarkers in saliva have been proposed for use in diagnosis of oral cancer, including metabolites, proteins, transcribed genes, miRNAs, genome alterations and epigenomic changes, as well as the microbiome. For the microbiome, however, saliva may not be optimal. Saliva bathes the entire oral cavity, resulting in a loss of information on the subsite specific composition of bacterial communities. Moreover, with saliva, there is no possibility to use each individual as his or her own control, and so account for the substantial variation in the oral microbiome amongst individuals. Non-invasively sampling the microbiome of oral lesions and corresponding normal tissue opens the possibility to not only detect cancer-associated changes at one time point, but the relative stability of the adult oral microbiome also offers the opportunity to monitor shifts in bacterial communities over time. Here we observed changes in the microbiome, which, in future larger studies, may be confirmed as a potential biomarker of oral cancers or pre-cancers, and may even have utility to discriminate patients with lymph node metastases. In addition, there are other challenges in clinical management of oral cancers and pre-cancers that would benefit from better diagnostic tools.

The actions of RvD1 in our model of hyperoxia-induced lung injury is consistent with results by Rogers et al., whereby increasing pup exposure to DHA via supplemented dams led to improvement in inflammation but no change in the characteristic alveolar simplification seen with hyperoxia. Similar to RvD1, LXA4 demonstrated a reduction in septal wall thickening with values similar to that of the lungs in healthy, normoxia mice. However, unique to LXA4 was the improvement in alveogenesis with a reduction in MLI and increase in RAC approaching those seen in the Room Air group. The selective change in gene expression induced by LXA4 that may account for this morphometric difference between the two groups is the increase in expression of TGFb2 and to a lesser degree, the increase in Smad3. It is possible that this change in TGFb2 at the gene and protein level is unrelated to the improvement of alveologenesis observed with LXA4. However, TGFb has been described to have an important role across the spectrum of alveolar development including preservation of normal alveologenesis, as described above. Whether an increase at the protein level of Smad3 occurs with LXA4 needs to be studied further to corroborate the trends observed in gene expression. In summary, this study identified several candidate pathways by which fatty acid derived terminal metabolites may attenuate hyperoxia-induced lung injury. These pathways include their wellestablished role in regulating inflammation, but also include novel pathways in modulating the extracellular matrix and activating TGFb2-Smad3. Additional interrogation of these pathways, across different doses and including identification of the cell types involved in these pathways, are important next steps. In addition, it is important to note that in adult models of acute lung injury other pathways of attenuation have been identified with the use of these agents emphasizing their pleiotropic effects. The current study has several important implications. This is the first paper to demonstrate the role and potential pathways by which long chain polyunsaturated fatty acid derived terminal metabolites ameliorate a common neonatal morbidity that is characterized by both dysregulated inflammation and altered organogenesis. Second, it begins to offer biologic plausibility to the clinical studies documenting a relationship between systemic DHA levels and the risk of BPD. Lastly, this is the first description of the role of RvD1 and LXA4 in modulating neonatal organogenesis. The deficits in systemic levels DHA and AA, as observed in the early postnatal period in the preterm infant, potentially would also lead to lower availability of RvD1 and LXA4. Thus, strategies to replete DHA and AA during the early postnatal period of the preterm infant and/or administration of RvD1 and LXA4 may represent potential therapeutic strategies to ameliorate the development of BPD.